LOCUS       sequence-159511-        6695 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6695)
  AUTHORS   Murphy LO, Smith S, Chen RH, Fingar DC, Blenis J
  TITLE     Molecular interpretation of ERK signal duration by immediate early 
            gene products.
  JOURNAL   Nat Cell Biol. 2002 Aug . 4(8):556-64.
  PUBMED    12134156
REFERENCE   2  (bases 1 to 6695)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Cell 
            Biol. 2002 Aug . 4(8):556-64."
COMMENT     Sequence Label: pcDNA3-FLAG-Fos WT
FEATURES             Location/Qualifiers
     source          1..6695
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    270..391
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(281..300)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     335..354
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(565..1359)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(676..695)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1286..1305
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1426..1755)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1440..1575
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1494..1513)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     1740..1760
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(1769..2197)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1879..1900)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2091..2110
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2243..2467
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2456..2473
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        2493..2511
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2493..2510
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(3786..3809)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3877..3896)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3878..3896)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3941..4144)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3970..3990)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4145..4524
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4696..4715
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4790..4894
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4895..4963,4964..5755)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5113..5132)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5926..6514
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6415..6434
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6668..6685
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct
      241 agctagaggt cgacggtata cagacatgat aagatacatt gatgagtttg gacaaaccac
      301 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
      361 tgtaaccatt ataagctgca ataaacaagt tggggtgggc gaagaactcc agcatgagat
      421 ccccgcgctg gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt
      481 catagaaggc ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg
      541 tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat
      601 gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc
      661 gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac
      721 acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg
      781 caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc gcgccttgag
      841 cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc
      901 gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc
      961 gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga
     1021 tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa
     1081 tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc
     1141 cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca gggcaccgga
     1201 caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc
     1261 atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc
     1321 ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt
     1381 ctcttgatca gatccgaaaa tggatataca agctcccggg agctttttgc aaaagcctag
     1441 gcctccaaaa aagcctcctc actacttctg gaatagctca gaggcagagg cggcctcggc
     1501 ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg
     1561 agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg
     1621 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga
     1681 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     1741 ccctaactga cacacattcc acagaattaa ttcgcgttaa atttttgtta aatcagctca
     1801 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     1861 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     1921 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     1981 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     2041 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     2101 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     2161 acacccgccg cgcttaatgc gccgctacag ggcgcgtggg gataccccct agagccccag
     2221 ctggttcttt ccgcctcaga agccatagag cccaccgcat ccccagcatg cctgctattg
     2281 tcttcccaat cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac
     2341 ctactcagac aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca
     2401 ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa
     2461 ggcacagtcg aggctgatca gcgagctcta gcatttaggt gacactatag aatagggccc
     2521 tctagatgca tgctcgagcg gccgccagtg tgatggatat ctgcagaatt cctcgaggga
     2581 agacgtgttt ctcctctctg taatgcacca gctcagtcag tgccggctgc cctgccttct
     2641 ctgactgctc acagggctag cagtgtgggc gagctcagtg agtcagagga gggctcgttg
     2701 ctgctgctgc cctttcggtg ggcagctgcg cagctaggga aggagtcagc ctcggggtag
     2761 gtgaagacaa aggaagacgt ataggtagtg cagctgggag tgcaggtgac aacgggagtg
     2821 cacaggggct ccagctctgt gaccatgggc cccatcccca gggaactgct gtgcagaggc
     2881 tcccagtctg ctgcatagaa ggaaccagac aggtccacat ctggcacaga gcgggcagtc
     2941 tccgagccac tgggcctaga tgatgccgga aacaagaagt catcaaaggg ttcagccttc
     3001 agctccatgt tgctaatgtt cttgaccggc tccaaggatg gcttgggctc agggtcattg
     3061 agaagaggca gggtgaaggc ctcctcagac tctggggtgg tagcctcagg cagaccccca
     3121 gtcaagtcca gggaggtcac agacatctcc tctgggaagc ccaggtcatt ggggatcttg
     3181 caggcaggtc ggtgggctgc caaaataaac tccagttttt ccttctcttt cagtagattg
     3241 gcaatctcgg tctgcaacgc agacttctcg tcttcaagtt gatctgtctc cgcttggagc
     3301 gtatctgtca gctccctcct ccgattccgg cacttggctg cagccatctt attcctttcc
     3361 cttcggattc tccgtttctc ttcctcttca ggagatagct gctctacttt gccccttctg
     3421 ccgatgctct gcgctctgcc gcctgacatg gtcttcacca ctcccgctct ggcgtaagcc
     3481 ccggtcgacg gggtggggag tccgtaagga tggggcgctc tggtctgcga tggggccacg
     3541 gaggagacca gagtgggctg caccagccac tgcaggtctg ggctggtgga gatggctgtc
     3601 accgtgggga taaagttggc actagagacg gacagatctg cgcaaaagtc ctgtgtgttg
     3661 acaggggagc ccatgctgga gaaggagtcg gctggggaat ggtagtagga aaggctgtcc
     3721 ccggccgggg aggcgctact gcagcgggag gatgacgcct cgtagtccgc gttgaaaccc
     3781 gagaacttgt cgtcatcgtc tttgtagtcc atggtgaatt ccagcacact ggcggccgtt
     3841 actagtggat ccgagctcgg taccaagctt gggtctccct atagtgagtc gtattaattt
     3901 cgataagcca gtaagcagtg ggttctctag ttagccagag agctctgctt atatagacct
     3961 cccaccgtac acgcctaccg cccatttgcg tcaatggggc ggagttgtta cgacattttg
     4021 gaaagtcccg ttgattttgg tgccaaaaca aactcccatt gacgtcaatg gggtggagac
     4081 ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt gatgtactgc caaaaccgca
     4141 tcaccatggt aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccata
     4201 aggtcatgta ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg
     4261 gcgtacttgg catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact
     4321 ccacccattg acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta
     4381 ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg cgggccattt accgtaagtt
     4441 atgtaacgcg gaactccata tatgggctat gaactaatga ccccgtaatt gattactatt
     4501 aataactagt caataatcaa tgtcaacgcg tatatctggc ccgtacatcg cgaagcagcg
     4561 caaaacgcct aaccctaagc agattcttca tgcaattgtc ggtcaagcct tgccttgttg
     4621 tagcttaaat tttgctcgcg cactactcag cgacctccaa cacacaagca gggagcagat
     4681 actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat aggggatcgg
     4741 gagatctccc gatccgtcga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4801 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     4861 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     4921 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     4981 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     5041 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     5101 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     5161 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     5221 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     5281 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     5341 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     5401 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     5461 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     5521 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     5581 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     5641 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     5701 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     5761 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5821 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5881 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5941 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     6001 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     6061 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     6121 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     6181 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     6241 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     6301 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6361 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6421 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6481 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6541 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     6601 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     6661 cgagcgcagc gagtcagtga gcgaggaagc ggaag
//
